title: Amniotic fluid stem cells morph into a cardiovascular lineage: analysis of a chemically induced cardiac and vascular commitment.
abstract: Mouse embryonic stem cells were previously observed along with mesenchymal stem cells from different sources, after being treated with a mixed ester of hyaluronan with butyric and retinoic acids, to show a significant increase in the yield of cardiogenic and vascular differentiated elements. The aim of the present study was to determine if stem cells derived from primitive fetal cells present in human amniotic fluid (hAFSCs) and cultured in the presence of a mixture of hyaluronic (HA), butyric (BU), and retinoic (RA) acids show a higher yield of differentiation toward the cardiovascular phenotype as compared with untreated cells. During the differentiation process elicited by exposure to HA + BU + RA, genes controlling pluripotency and plasticity of stem cells, such as Sox2, Nanog, and Oct4, were significantly downregulated at the transcriptional level. At this point, a significant increase in expression of genes controlling the appearance of cardiogenic and vascular lineages in HA + BU + RA-treated cells was observed. The protein expression levels typical of cardiac and vascular phenotypes, evaluated by Western blotting, immunofluorescence, and flow cytometry, were higher in hAFSCs cultured in the presence of HA + BU + RA, as compared with untreated control cells. Appearance of the cardiac phenotype was further inferred by ultrastructural analysis using transmission and scanning electron microscopy. These results demonstrate that a mixture of HA + BU + RA significantly increased the yield of elements committed toward cardiac and vascular phenotypes, confirming what we have previously observed in other cellular types.
authors.affiliation: Department of Biomedical sciences, University of Sassari, Sassari, Italy ; Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Bologna, Italy ; Department of Regenerative Medicine, Rinaldi Fontani Institute, Florence, Italy.
authors.forename: Margherita
authors.initials: M
authors.lastname: Maioli
authors.forename: Giovanni
authors.initials: G
authors.lastname: Contini
authors.forename: Sara
authors.initials: S
authors.lastname: Santaniello
authors.forename: Pasquale
authors.initials: P
authors.lastname: Bandiera
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Pigliaru
authors.forename: Raimonda
authors.initials: R
authors.lastname: Sanna
authors.forename: Salvatore
authors.initials: S
authors.lastname: Rinaldi
authors.forename: Alessandro P
authors.initials: AP
authors.lastname: Delitala
authors.forename: Andrea
authors.initials: A
authors.lastname: Montella
authors.forename: Luigi
authors.initials: L
authors.lastname: Bagella
authors.forename: Carlo
authors.initials: C
authors.lastname: Ventura
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D001808:Blood Vessels / Q000166:cytology; D015153:Blotting, Western; D020148:Butyric Acid / Q000737:chemistry / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D015536:Down-Regulation / Q000187:drug effects*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D018398:Homeodomain Proteins / Q000235:genetics; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry / Q000494:pharmacology; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D009206:Myocardium / Q000166:cytology; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D010641:Phenotype; D055748:SOXB1 Transcription Factors / Q000235:genetics; D013234:Stem Cells / Q000378:metabolism*; D014212:Tretinoin / Q000737:chemistry / Q000494:pharmacology
chemical_list: D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D020148:Butyric Acid; D014212:Tretinoin; D006820:Hyaluronic Acid
keywords: GATA-4; Nkx-2.5; butyric acid; fetal cells; human amniotic fluid; hyaluronic acid; prodynorphin; retinoic acid; von Willebrand factor